Role of the renal organic anion transporter OAT1 in metabolism and physiology

项目摘要/摘要 由于新的监管指南，有机阴离子转运体(OAT1)--一种被PI发现的转运体 小组(作为NKT)，并参与消除许多常见药物--收到了相当大的 请注意。但几乎所有已发表的研究都继续关注普通药物，而不是内源性药物 转运蛋白的底物，从进化的角度来看是高度保守的。代谢组学 对我们的Oat1基因敲除小鼠(Oat1KO)的分析以及体外转运数据表明，OAT1是一种 代谢产物、信号分子、维生素、肠道微生物组产品的主要运输者，以及 抗氧化剂。我们先前对Oat1KO改变的代谢的基因组规模的代谢重建 正常饮食条件下的小鼠使我们能够创造出第一个多特异性的药物转运蛋白(OAT1)- 以代谢网络为中心--这进一步得到了相当大的代谢组学和体外数据的支持。 许多途径(例如，脂肪酸、肠道微生物组产物、支链氨基酸)在很大程度上 已证实的~150-200代谢物网络在代谢性疾病中也是重要的，例如 糖尿病、代谢综合征、肥胖症和肠道微生物群相关疾病。在此提案中：1)我们将使用 Oat1 KO小鼠的饮食调整(如高脂肪、支链氨基酸)和抗生素治疗 确定燕麦1在适用于代谢性疾病的条件下的作用。我们将测试绑定 代谢组学揭示了不同饮食条件(或疾病模型)下的代谢产物 吞吐量未标记(BLI)分析，以量化代谢物与OAT1的相互作用；这一信息将 用于区分代谢产物的优先顺序，以便进行后续的摄取分析，包括使用放射性标记化合物。2) 然后，我们将使用这些信息，以及来自不同基因敲除组织的基因表达数据 饮食条件，以重建每个这些饮食(病理模型)条件的代谢网络- -使用我们以前成功使用的方法，在以下条件下创建了以OAT1为中心的代谢网络 正常的饮食条件。总之，这些研究将有助于确定OAT1在调节中的独特作用 在这些失调的代谢状态下的异常新陈代谢。最后，我们将讨论这些信息将如何 为了解服用OAT1-1患者的药物诱导代谢综合征的各个方面奠定基础 运输毒品。我们在必要的技术方面拥有成熟的专业知识和一支世界级的团队 确保成功的合作者。

PROJECT SUMMARY/ABSTRACT Owing to new regulatory guidelines, the organic anion transporter (OAT1)--a transporter discovered by the PI's group (as NKT) and which is involved in the elimination of many common drugs--has received considerable attention. But nearly all the published studies continue to focus on common drugs rather than endogenous substrates of the transporter, which is highly conserved from an evolutionary standpoint. Metabolomics analysis of our Oat1 knockout mice (Oat1KO), as well as in vitro transport data, indicate that OAT1 is a major transporter of metabolites, signaling molecules, vitamins, gut microbiome products, and antioxidants. Our previous genome-scale metabolic reconstructions of altered metabolism of the Oat1KO mouse under normal dietary conditions enabled us to create the first multispecific "drug" transporter (OAT1)- centered metabolic network--which was further supported by considerable metabolomics and in vitro data. Many pathways (e.g., fatty acids, gut microbiome products, branched chain amino acids) in this largely validated network of ~150-200 metabolites are also known to be important in metabolic diseases such as diabetes, metabolic syndrome, obesity, and gut microbiome-associated illness. In this proposal: 1) We will use dietary modifications (e.g., high fat, branched chain amino acids) and antibiotic treatment of the Oat1 KO mice to define the role of Oat1 under conditions applicable to metabolic diseases. We will test the binding of metabolites revealed by metabolomics under the various dietary conditions (or disease models) using a high throughput unlabeled (BLI) assay to quantify the interaction of metabolites with OAT1; this information will be used to prioritize metabolites for subsequent uptake assays involving the use of radiolabeled compounds. 2) We will then use this information, together with gene expression data from the knockout tissues under various dietary conditions, to reconstruct a metabolic network for each of these dietary (pathological model) conditions- -using methods we have previously successfully used to create an "OAT1-centered metabolic network" under normal dietary conditions. Together, these studies will help define the unique roles of OAT1 in regulation of aberrant metabolism in these dysregulated metabolic states. Finally, we discuss how this information will set the stage for understanding aspects of drug-induced metabolic syndrome in patients taking OAT1- transported drugs. We have proven expertise in the necessary techniques and a team of world-class collaborators to ensure success.